COI metabarcoding from ASU sample
Samples were amplified using the newly designed degenerate primer set targeting a 313-bp fragment of cytochrome c oxidase subunit 1 (COI) (forward IIICRrev: GGNTGAACNGTNTAYCCNCC; reverse HBR2d: TAWACTTCDGGRTGNCCRAARAAYCA) that has been developed to amplify a wide range of phyla across eukaryotes and algae. Forward and reverse primers were indexed with 1113 nucleotide long tags differentiated by at least three different nucleotides (for details see: Corse et al., 2017), and a 0 to 3 nt heterogeneity spacer (none/N/NN/NNN) was added to mitigate the issues caused by low sequence diversity in Illumina amplicon sequencing (Fadrosh et al., 2014). A unique combination of tags was used for each sample. Each 15-L amplification mix consisted of 7.5-L QIAGEN Multiplex Master Mix, 2.5-L QIAGEN Nuclease-Free H20, 1.5 L of each respective tagged primer (2 M) and 2-L DNA